Multi-substituted imidazolines and method of use thereof

ABSTRACT

A new class of imidazolines as 4-position acids or esters with very potent anti-inflammatory as well as antimicrobial activity is described. The synthesis of these imidazolines includes a multicomponent reaction applicable to a combinatorial synthetic approach. The combination of these two key characteristics provides an effective therapeutic drug in the treatment of septic shock as well as many other inflammatory (arthritis and asthma) and infectious disorders. The use of this novel class of non-steroidal agents as anti-inflammatory agents (for the treatment of asthma etc.), antibacterial agents and antiseptic agents is described. The compounds are also useful in the treatment of tumors (such as cancers). The imidazolines are potent inhibitors of the transcription factor NF-κB as well as potent activity against the Gram (+) bacterium  B. subtillus  and  B. cereus  with MIC values in the range of 50 μm/mL.

CROSS REFERENCE TO RELATED APPLICATION

This application is a divisional of U.S. patent application Ser. No.10/347,323 filed on Jan. 17, 2003, now U.S. Pat. No. 6,878,735, whichclaims priority to U.S. Provisional Patent Application Ser. No.60/385,162 filed on May 31, 2002.

BACKGROUND OF THE INVENTION

(1) Field of the Invention

The present invention relates to novel multi-substituted 4-acid or alkylester or amide imidazolines and to a process for their preparation. Inparticular the present invention relates to the multi-substitutedimidazolines containing a 4-acid or an ester group which inhibit NFκB orNFκB kinase, are anti-inflammatory and/or antimicrobial and/orchemopotentiator and/or chemosensitizers of anticancer agents.

(2) Description of Related Art

Chronic airway inflammation as seen with asthma, is associated with theover expression of inflammatory proteins called cytokines. In addition,other inflammatory mediators, such as IL-1 and TNF, play a major role injoint diseases such as rheumatoid arthritis. All of these inflammatoryproteins are highly regulated by the nuclear transcription factor kappaB (NF-κB) (Yamamoto, Y., et al., J. Clin Invest 107 135-142 (2001); andHart, L. A., et al., Am J Respir Crit Care Med 158 1585-1592 (1998)).Inhibition of this regulatory protein or its kinase by anti-inflammatorydrugs has been shown to be effective in the treatment of these diseases(Yamamoto, Y., et al., J. Clin Invest 107 135-142 (2001); Coward, W. R.,et al., Clin Exp Allergy 28 Suppl 3, 42-46 (1998); Badger, A. M., etal., J. Pharmacol Exp Ther 290 587-593 (1999); Breton, J. J., et al., JPharmacol Exp Ther 282 459-466 (1997); Roshak, A., et al., J PharmacolExp Ther 283 955-961 (1997); Kopp, E., et al., Science 265 956-959(1994); Ichiyama, T., et al., Brain Res 911 56-61 (2001); Hehner, S. P.,et al., J Immunol 163 5617-5623 (1999); Natarajan, K., et al., Proc NatlAcad Sci USA 93 9090-9095 (1996); and Fung-Leung, W. P., et al.,Transplantation 60 362-368 (1995)). The common anti-inflammatory agent,aspirin, and aspirin-like drugs, the salicylates, are widely prescribedagents to treat inflammation and their effectiveness has been attributedto NF-κB inhibition. However, in order to treat chronic inflammations,the cellular levels of these salicylates need to be at very highconcentration and are generally prescribed at 1-3 miliMolar plasmaconcentrations (Science 265, 956-959 (1994)).

Since the discovery of penicillin, over 100 antibacterial agents havebeen developed to combat a wide variety of bacterial infections. Today,the clinically used antibacterial agents mainly consists of β-lactams(penicillins, carbapenems and cephalosporins), aminoglycosides,tetracyclines, sulfonamides, macrolides (erythromycin), quinolones, andthe drug of last resort: vancomycin (a glycopeptide). In recent years,many new strains of bacteria have developed resistance to these drugsthroughout the world. There is a need for new antimicrobials.

There is considerable interest in modulating the efficacy of currentlyused antiproliferative agents to increase the rates and duration ofantitumor effects associated with conventional antineoplastic agents.Conventional antiproliferative agents used in the treatment of cancerare broadly grouped as chemical compounds which (1) affect the integrityof nucleic acid polymers by binding, alkylating, inducing strand breaks,intercalating between base pairs or affecting enzymes which maintain theintegrity and function of DNA and RNA; and (2) chemical agents that bindto proteins to inhibit enzymatic action (e.g. antimetabolites) or thefunction of structural proteins necessary for cellular integrity (e.g.antitubulin agents). Other chemical compounds that have been identifiedto be useful in the treatment of some cancers include drugs which blocksteroid hormone action for the treatment of breast and prostate cancer,photochemically activated agents, radiation sensitizers and protectors.

Of special interest to this invention are those compounds that directlyaffect the integrity of the genetic structure of the cancer cells.Nucleic acid polymers such as DNA and RNA are prime targets foranticancer drugs. Alkylating agents such as nitrogen mustards,nitrosoureas, aziridine (such as mitomycin C) containing compoundsdirectly attack DNA. Metal coordination compounds such as cisplatin andcarboplatin similarly directly attack the nucleic acid structureresulting in lesions that are difficult for the cells to repair, which,in turn, can result in cell death. Other nucleic acid affectingcompounds include anthracycline molecules such as doxorubicin, whichintercalates between the nucleic acid base pairs of DNA polymers,bleomycin which causes nucleic acid strand breaks, fraudulentnucleosides such as pyrimidine and purine nucleoside analogs which areinappropriately incorporated into nucleic polymer structures andultimately cause premature DNA chain termination. Certain enzymes thataffect the integrity and functionality of the genome can also beinhibited in cancer cells by specific chemical agents and result incancer cell death. These include enzymes that affect ribonucleotidereductase (.e.g. hydroxyurea, gemcitabine), topoisomerase I (e.g.camptothecin) and topoisomerase II (e.g. etoposide).

The topoisomerase enzymes affect the structure of supercoiled DNA,because most of the functions of DNA require untwisting. Topoisomerase I(top 1) untwists supercoiled DNA, breaking only one of the two strands,whereas topoisomerase II (top 2) breaks both.

Topoisomerase I inhibition has become important in cancer chemotherapythrough the finding that camptothecin (CPT), an alkaloid of plantorigin, is the best known inhibitor of top 1 and is a very potentanticancer agent. CPT is contained in a Chinese tree, Camptothecaacuminata. A number of analogs have become approved for commercial useto treat a number of tumor types. These include CPT-11 (irinotecan) andtopotecan.

While the clinical activity of camptothecins against a number of typesof cancers are demonstratable, improvements in tumor response rates,duration of response and ultimately patient survival are still sought.The invention described herein demonstrates the novel use which canpotentiate the antitumor effects of chemotherapeutic drugs, includingtopoisomerase I inhibitors, in particular, camptothecins.

Relevant Literature

Cancer Chemotherapeutic Agents, W. O. Foye, ed., (ACS, Washington, D.C.)(1995)); Cancer Chemotherapy Handbook, R. T. Dorr and D. D. VonHoff,(Appleton and Lange, Norwalk, Conn.) (1994); and M. P. Boland,Biochemical Society Transactions (2001) volume 29, part 6, p 674-678.DNA damage signaling and NF-κB: implications for survival and death inmammalian cells.

Invasive infection with Gram positive or Gram negative bacteria oftenresults in septic shock and death. Invasion of the blood stream by bothtypes of bacteria (Gram positive and Gram negative) causes sepsissyndrome in humans as a result of an endotoxin, Lipopolysaccharide (LPS)(H. Bohrer, J. Clin. Invest. 972-985 (1997)), that triggers a massiveinflammation response in the host. The mechanism by which LPS causedseptic shock is through the activation of the transcription factorNF-κB. Activation of this protein by its kinase initiates the massiverelease of cytokines resulting in a potentially fatal septic shock. Forexample, the pneumococcus bacteria is the leading cause of death with amortality rate of 40% in otherwise healthy elderly individuals andstaphylococcal infections are the major cause of bacteremia in UShospitals today. Septic shock, caused by an exaggerated host response tothese endotoxins often leads to multiple organ dysfunction, multipleorgan failure, and remains the leading cause of death in traumapatients.

NF-κB has been indicated to inhibit apoptosis (programmed cell death).Many clinically used chemotherapeutic agents (including the vincaalkaloids, vincristine and vinblastinc, camptothecin and many others)have recently been shown to activate NF-κB resulting in a retardation oftheir cytotoxicity. This form of resistance is commonly referred to asNF-κB mediated chemoresistance. Inhibition of NF-κB has shown toincrease the sensitivity to chemotherapeutic agents of tumor cells andsolid tumors.

REFERENCES

Cusack, J. C.; Liu, F.; Baldwin, A. S. NF-kappa B and chemoresistance:potentiation of cancer drugs via inhibition of NF-kappa B. Drug ResistUpdat 1999, 2, 271-273, Mayo, M. W.; Baldwin, A. S. The transcriptionfactor NF-kaapaB: control of oncogenesis and cancer therapy resistance,Biochim Biophys Acta 2000, 1470, M55-62. Wang, C. Y.; Mayo, M. W.;Baldwin, A. S., Jr. TNF- and cancer therapy-induced apoptosis;potentiation by inhibition of NF-kappaB. Science 1996, 274, 784-787.(Cusack, J. C., Jr.; Liu, R.; Baldwin, A. S., Jr. Induciblechemoresistance to7-ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxycamptothecin(CPT-11) in colorectal cancer cells and a xenograft model is overcome byinhibition of nuclear factor-kaapaB activation. Cancer Res 2000, 60,2323-2330. Brandes, L. M.; Lin, Z. P.; Patierno, S. R.; Kennedy, K. A.Reversal of physiological stress-induced resistance to topoisomerase IIinhibitors using an inducible phosphorylation site-deficient mutant of 1kappa B alpha. Mol Pharmacol 2001, 60, 559-567, Arlt, A.; Vorndamm, J.;Breitenbroich, M.; Folsch, U. R.; Kalthoff, H. et al. Inhibition ofNF-kappaB sensitizes human pancreatic carcinoma cells to apoptosisinduced by etoposide (VP16) or doxorubicin. Oncogene 2001, 20, 859-868.Cusack, J. C., Jr.; Liu, R.; Houston, M.; Abendroth, K.; Elliott, P. J.et al. Enhanced chemosensitivity to CPT-11 with proteasome inhibitorPS-341; implications for systemic nuclear factor kappaB inhibition.Cancer Res 2001 61, 3535-3540.

1,3 Dipolar cycloadditions reactions utilizing azlactones of “munchones”provide a general route for the synthesis of pyrroles and imidazoles(Hershenson, F. M. P., Synthesis 999-1001 (1988); Consonni, R. C., etal., J. chem. Research (S) 188-189 (1991); and Bilodeau, M. T. C., J.Org. Chem. 63 2800-2801 (1998)). This approach has not yet been reportedfor the imidazoline class of heterocycles. The synthetic andpharmacological interest in efficient syntheses of imidazolines hasfueled the development of several diverse synthetic approaches(Puntener, K., et al., J. Org Chem 65 8301-8306 (2000); Hsiao, Y. H., J.Org. Chem. 62 3586-3591 (1997)). Recently, Arndtsen et al reportedsynthesis of symmetrically substituted imidazoline-4-carboxylic acidsvia a Pd-catalyzed coupling of an imine, acid chloride and carbonmonoxide (Dghaym, R. D. D., et al., Angew. Chem. Int. Ed. Engl. 403228-3230 (2001)). In addition, diastereoselective 1,3-dipolarcycloaddition of azomethine ylides has been reported from amino acidesters with enantiopure sulfinimines to yield Nsulfinyl imidazolidines(Viso, A., et al., J. Org. Chem. 62 2316-2317 (1997)).

U.S. Pat. No. 6,318,978 to Ritzeler et al describes 3,4-benzimidazoleswhich are structurally quite different than those of the presentinvention. They inhibit NFkB kinase. As can be seen, activity isretained where there are numerous different substituents in theimidazoline and benzene rings. M. Karin, Nature immunology, 3, 221-227(2002); Baldwin, J. Clin. Invest., 3, 241-246 (2001); T. Huang et al, J.Biol. Chem., 275, 9501-9509 (2000); and J. Cusack and Baldwin, CancerResearch, 60, 2323-2330 (2000) describe the effect of activation of NFkBon cancer. U.S. Pat. Nos. 5,804,374 and 6,410,516 to Baltimore describeNFkB inhibition which are incorporated by reference.

Patents of interest for the general methodology of inhibition are setforth in U.S. Pat. No. 5,821,072 to Schwartz et al and U.S. Pat. No.6,001,563 to Deely et al.

OBJECTS

It is an object of the present invention to provide novel compoundswhich are anti-inflammatory, antimicrobial and inhibit NFκB or NFκBkinase. It is also an object of the present invention to provide forinhibition of cancers by inhibition of chemoresistance. It is further anobject of the present invention to provide a novel process for thepreparation of such compounds. These and other objects will becomeincreasingly apparent by reference to the following description and thedrawings.

SUMMARY OF THE INVENTION

The present invention relates to a method for inhibiting inflammation ina mammal which comprises administering a multi-substituted 4-acid or4-alkyl ester imidazoline to the mammal in an amount sufficient toinhibit the inflammation.

The present invention also relates to a method of inhibiting theactivation of the NF-κB protein by inhibition of the degradation of theinhibitory protein, I kappa B, or its kinases and the ability to inhibitNF-κB which comprises of contacting the protein or its activatingproteins with a multi-substituted 4-acid or 4-alkyl ester or amideimidazoline in an amount sufficient to inhibit activation of theprotein.

The present invention further relates to a method of inhibiting a cancerwhich comprises contacting the cancer with a multi-substitutedimidazoline in an amount sufficient to inhibit the cancer.

The present invention relates to an imidazoline of the formula:

wherein R₁, R₂, R₃ and R₄ are selected from the group consisting ofalkyl, acyl, aryl, arylalkyl, heteroaryl containing 5 to 14 ringmembers, and heterocyclic containing 5 to 12 ring members; X is selectedfrom the group consisting of O and S; and R₅ is selected from the groupconsisting of hydrogen, alkyl, acyl, aryl arylalkyl, heteroaryl, NH₂,NH—R₆ and

where R₆ and R₇ are selected from the group consisting of hydrogen,alkyl, aryl, arylalkyl, and heteroaryl and heterocyclic, which may bethe same or different.

Further the present invention relates to an imidazoline of the formula

wherein R₁, R₂, R₃ and R₄ are selected from the group consisting ofalkyl, acyl, aryl, arylalkyl, heteroaryl containing 5 to 14 ringmembers, and heterocyclic containing 5 to 12 ring members; and whereinR₈ and R₉ and selected from the group consisting of hydrogen, alkyl,aryl, arylalkyl, heteroaryl and heterocyclic, which may be the same ordifferent.

Further, the present invention relates to a process for the preparationof an amino imidazoline which comprises reacting an imidazoline of theformula:

wherein R₁, R₂, R₃ and R₄ are selected from the group consisting ofalkyl, acyl, aryl, arylalkyl, heteroaryl containing 5 to 14 ringmembers, and heterocyclic containing 5 to 12 ring members; X is selectedfrom the group consisting of O and S; and R₅ is selected from the groupconsisting of hydrogen, alkyl, acyl, aryl arylalkyl, heteroaryl, NH₂,NH—R₆ and

where R₆ and R₇ and selected from the group consisting of hydrogen,alkyl, aryl, arylalkyl, and heteroaryl and heterocyclic, which may bethe same or different, with an amine of the formula:

to produce a compound of the formula:

wherein R₈ and R₉ are selected from the group consisting of hydrogen,alkyl, acyl, arylalkyl and heteroalkyl, which may be the same ordifferent.

The present invention relates to an imidazoline of the formula

wherein R₁, R₂, R₃ and R₄ are selected from the group consisting ofalkyl, acyl, aryl, aralkyl, heteroaryl containing 5 to 14 ring members,and heterocyclic containing 5 to 12 ring members; and R₅ is selectedfrom the group consisting of hydrogen and an alkyl group, all of whichare optionally substituted.

The present invention particularly relates to an imidazoline of theformula:

wherein R₁, R₂, R₃ and R₄ are each individually selected from the groupconsisting of alkyl, acyl, aryl, arylalkyl, heteroaryl containing 5 to14 ring members, and heterocyclic containing 5 to 12 ring members; andR₅ is selected from the group consisting of hydrogen and an alkyl group,all of which are optionally substituted. Preferably R₁ is phenyl; R₄ isbenzyl; R₅ is lower alkyl containing 1 to 4 carbon atoms. Alsopreferably R₅ is ethyl; R₂ is lower alkyl containing 1 to 4 carbonatoms. Most preferably R₂ is methyl and R₃ is selected from the groupconsisting of phenyl and substituted phenyl.

The imidazoline(Compound 1) wherein R₁ is phenyl, R₂ is methyl, R₃ isphenyl, R₄ is benzyl and R₅ is H is a preferred compound. Theimidazoline (Compound 2) wherein R₁ is phenyl, R₂ is methyl, R₃ is4-methoxyphenyl, R₄ is benzyl and R₅ is H is a preferred compound. Theimidazoline (Compound 3) wherein R₁ is phenyl, R₂ is methyl, R₃ isphenyl, R₄ is 4-fluorophenyl and R₅ is H is a preferred compound. Theimidazoline (compound 4) wherein R₁ is phenyl, R₂ is phenyl, R₃ isphenyl, R₄ is benzyl and R₅ is H is a preferred compound. Theimidazoline (Compound 5) wherein R₁ is phenyl, R₂ is1H-indol-3-ylmethyl, R₃ is phenyl, R₄ is benzyl and R₅ is H is apreferred compound. The imidazoline (Compound 6) wherein R₁ is phenyl,R₂ is methyl, R₃ is pyridin-4-yl, R₄ is benzyl and R₅ is H is apreferred compound. The imidazoline (Compound 7) wherein R₁ is phenyl,R₂ is methyl, R₃ is phenyl, R₄ is H and R₅ is H is a preferred compound.The imidazoline (Compound 8) wherein R₁ is phenyl, R₂ is methyl, R₃ isethoxycarbonyl, R₄ is H and R₅ is H is a preferred compound. Theimidazoline (Compound 9) wherein R₁ is phenyl, R₂ is methyl, R₃ ispyridin-4-yl, R₄ is benzyl and R₅ is Et is a preferred compound. Theimidazoline (Compound 10) wherein R₁ is phenyl, R₂ is methyl, R₃ isphenyl, R₄ is benzyl and R₅ is Et is a preferred compound.

The present invention also relates to a process for the preparation ofimidazoline of the formula:

wherein R₁, R₂, R₃ and R₄ are selected from the group consisting ofalkyl, acyl, aryl, aralkyl, heteroaryl containing 5 to 14 ring members,and heterocyclic containing 5 to 12 ring members; and R₅ is selectedfrom the group consisting of hydrogen and an alkyl group, all of whichare optionally substituted, which comprises:

(a) reacting a reaction mixture of

-   -   (1) an oxazolone of the formula:

-   -   (2) a ketone of the formula:        R₃═O; and    -   (3) an amine of the formula:        H₂N—R₄        in the presence of trimethyl silyl chloride or an acid chloride        and a solvent for the reactants in the absence of water in the        presence of a non-reactive gas and at a temperature between        about 0 and 100° C. to produce the imidazoline; and

(b) separating the imidazoline from the reaction mixture. Theimidazoline can be esterified by reaction with an alcohol. Theimidazoline is most preferably esterified by reaction with the alcoholand sulfonyl dichloride.

The present invention relates to a method for inhibiting inflammation ina mammal which comprises administering an imidazoline of the formula:

wherein R₁, R₂, R₃ and R₄ are selected from the group consisting ofalkyl, acyl, aryl, aralkyl, heteroaryl containing 5 to 14 ring members,and heterocyclic containing 5 to 12 ring members; and R₅ is selectedfrom the group consisting of hydrogen and an alkyl group, all of whichare optionally substituted, to the mammal in an amount sufficient toinhibit the inflammation. Preferably the mammal is human. The mammal canbe a lower mammal. The administration can be oral, topical, or byinjection (such as intravenous) into the mammal.

The present invention also relates to a method for inhibiting amicroorganism which comprises:

administering an effective amount of a compound of the formula:

wherein R₁, R₂, R₃ and R₄ are selected from the group consisting ofalkyl, acyl, aryl, aralkyl, heteroaryl containing 5 to 14 ring members,and heterocyclic containing 5 to 12 ring members; and R₅ is selectedfrom the group consisting of hydrogen and an alkyl group, all of whichare optionally substituted, to inhibit the microorganism. The inhibitioncan be in vitro or in vivo. The administration can be to a lower mammalor to a human. The administration can be oral, by injection into themammal, or topical.

Further, the present invention relates to a method of inhibitingdegradation of a protein which is NF-κB or NF-κB kinase which comprisescontacting the protein with a compound of the formula:

wherein R₁, R₂, R₃ and R₄ are selected from the group consisting ofalkyl, acyl, aryl, aralkyl, heteroaryl containing 5 to 14 ring members,and heterocyclic containing 5 to 12 ring members; and R₅ is selectedfrom the group consisting of hydrogen and an alkyl group, all of whichare optionally substituted. The compounds are also useful in thetreatment of tumors (cancers) where NFkB is involved. The inhibition ispreferably in vivo.

R₁ is

-   (1) phenyl, mono- or disubstituted independently of one another by-   (1)(1)—CN;-   (1)(2)—NO₂;-   (1)(3)—O—(C₁-C₄)-alkyl;-   (1)(4)—NH₂; or-   (1)(5)—(C₁-C₄)-alkyl-NH₂;-   (1)(6)—x, wherein x is a halogen.-   (2) heteroaryl having 5 to 14 ring members, in which the heteroaryl    is unsubstituted or mono-, di-, or trisubstituted independently of    one another by —N—R¹⁴, in which R¹⁴ is —(C₁-C₆)-alkyl,    —(C₃-C₆)-cycloalkyl, phenyl, halogen, —OH, or —(C₁-C₄)-alkyl; or-   (3) a heterocycle having 5 to 12 ring members, in which the    heterocycle is unsubstituted or mono-, di-, or trisubstituted    independently of one another by —N—R¹⁴, in which R¹⁴ is    —(C₁-C₆)-alkyl, —(C₃-C₆)-cycloalkyl, phenyl, halogen, —OH, or    —(C₁-C₄)-alkyl.

The term “halogen” is understood as meaning fluorine, chlorine, bromine,or iodine. The term “aryl” is understood as meaning aromatic hydrocarbongroups having 6 to 14 carbon atoms in the ring. (C₆-C₁₄)-Aryl groupsare, for example, phenyl, naphthyl, for example, 1-naphthyl, 2-naphthyl,biphenylyl, for example, 2-biphenylyl, 3-biphenylyl, and 4-biphenylyl,anthryl, or fluorenyl. Biphenylyl groups, naphthyl groups, and, inparticular, phenyl groups are preferred aryl groups. Aryl groups, inparticular phenyl groups, can be mono-substituted or polysubstituted,preferably monosubstituted, disubstituted, or trisubstituted, byidentical or different groups, preferably by groups selected from(C₁-C₈)-alkyl, in particular (C₁-C₄)-alkyl, (C₁-C₈)-alkoxy, inparticular (C₁-C₄)-alkoxy, halogen, nitro, amino, trifluoromethyl,hydroxyl, hydroxy-(C₁-C₄)-alkyl such as hydroxymethyl, 1-hydroxyethyl,or 2-hydroxyethyl, methylenedioxy, ethylenedioxy, formyl, acetyl, cyano,hydroxycarbonyl, aminocarbonyl, (C₁-C₄)-alkoxycarbonyl, phenyl, phenoxy,benzyl, benzyloxy, or tetrazolyl. Further, when aryl is phenyl, phenylis optionally mono- or disubstituted independently of one another by—CN, —NO₂, —O—(C₁-C₄)-alkyl, —N(R¹¹)₂, —NH—C(O)—R¹¹, —S(O)_(x)R¹, inwhich x is the integer 0, 1, or 2, —C(O)—R¹¹, in which R¹¹ is as definedabove, or —(C₁-C₄)-alkyl-NH₂. The same applies, for example, to groupssuch as arylalkyl or arylcarbonyl. Arylalkyl groups are, in particular,benzyl and also 1- and 2-naphthylmethyl, 2-, 3-, and 4-biphenylylmethyl,and 9-fluorenylmethyl. Substituted arylalkyl groups are, for example,benzyl groups and naphthylmethyl groups substituted in the aryl moietyby one or more (C₁-C₈)-alkyl groups, in particular (C₁-C₄)-alkyl groups,for example, 2-, 3-, and 4-methylbenzyl, 4-isobutylbenzyl,4-tert-butylbenzyl, 4-octylbenzyl, 3,5-dimethylbenzyl,pentamethylbenzyl, 2-, 3-, 4-, 5-, 6-, 7-, and8-methyl-1-naphthylmethyl, 1-, 3-, 4-, 5-, 6-, 7-, and8-methyl-2-naphthylmethyl, by one or more (C₁-C₈)-alkoxy groups, inparticular (C₁-C₄)-alkoxy groups, benzyl groups, and naphthylmethylgroups substituted in the aryl moiety for example, 4-methoxybenzyl,4-neopentyloxybenzyl, 3,5-dimethoxybenzyl, 3,4-methylenedioxybenzyl,2,3,4-trimethoxybenzyl, nitrobenzyl groups, for example, 2-, 3-, and4-nitrobenzyl, halobenzyl groups, for example, 2-, 3-, and 4-chloro- and2-, 3-, and 4-fluorobenzyl, 3,4-dichlorobenzyl, pentafluorobenzyl,trifluoromethylbenzyl groups, for example, 3- and4-trifluoromethylbenzyl, or 3,5-bis(trifluoromethyl)benzyl.

In monosubstituted phenyl groups, the substituent can be located in the2-position, the 3-position, or the 4-position. Disubstituted phenyl canbe substituted in the 2,3-position, the 2,4-position, the 2,5-position,the 2,6-position, the 3,4-position, or the 3,5-position. Intrisubstituted phenyl groups, the substituents can be located in the2,3,4-position, the 2,3,5-position, the 2,4,5-position, the2,4,6-position, the 2,3,6-position, or the 3,4,5-position.

The explanations for the aryl groups apply accordingly to divalentarylene groups, for example, to phenylene groups that can be present,for example, as 1,4-phenylene or as 1,3-phenylene.

Phenylene-(C₁-C₆)-alkyl is in particular phenylenemethyl(—C₆H₄—CH₂—) andphenyleneethyl. (C₁-C₆). Alkylenephenyl is in particular methylenephenyl(—CH₂—C₆H₄—). Phenylene-(C₁-C₆)-alkenyl is in particularphenyleneethenyl and phenylenepropenyl.

The expression “heteroaryl having 5 to 14 ring members” represents agroup of a monocyclic or polycyclic aromatic system having 5 to 14 ringmembers, which contains 1, 2, 3, 4, or 5 heteroatoms as ring members.Examples of heteroatoms are N, O, and S. If a number of heteroatoms arecontained, these can be identical or different. Heteroaryl groups canlikewise be monosubstituted or polysubstituted, preferablymonosubstituted, disubstituted, or trisubstituted, by identical ordifferent groups selected from (C₁-C₈)-alkyl, in particular(C₁-C₄)-alkyl, (C₁-C₈)-alkoxy, in particular (C₁-C₄)-alkoxy, halogen,nitro, —N(R¹¹)₂, trifluoromethyl, hydroxyl, hydroxy-(C₁-C₄)-alkyl suchas hydroxymethyl, 1-hydroxyethyl, or 2-hydroxyethyl, methylenedioxy,formyl, acetyl, cyano, hydroxycarbonyl, aminocarbonyl,(C₁-C₄)-alkoxycarbonyl, phenyl, phenoxy, benzyl, benzyloxy, ortetrazolyl. Heteroaryl having 5 to 14 ring members preferably representsa monocyclic or bicyclic aromatic group which contains 1,2,3, or 4, inparticular 1, 2, or 3, identical or different heteroatoms selected fromN, O, and S, and which can be substituted by 1,2,3, or 4, in particular1, 2, or 3, identical or different substituents selected from(C₁-C₆)-alkyl, (C₁-C₆)-alkoxy, fluorine, chlorine, nitro, —N(R¹¹)₂,trifluoromethyl, hydroxyl, hydroxy(C₁-C₄)-alkyl, (C₁-C₄)-alkoxycarbonyl,phenyl, phenoxy, benzyloxy, and benzyl. Heteroaryl particularlypreferably represents a monocyclic or bicyclic aromatic group having 5to 10 ring members, in particular a 5-membered or 6-membered monocyclicaromatic group which contains 1, 2, or 3, in particular 1 or 2,identical or different heteroatoms selected from N, O, and S, and can besubstituted by 1 or 2 identical or different substituents selected from(C₁-C₄)-alkyl, halogen, hydroxyl, —N(R¹¹)₂, (C₁-C₄)-alkoxy, phenyl,phenoxy, benzyloxy, and benzyl. R¹¹ is as defined in substituent R⁹ offormula I.

The expression “heterocycle having 5 to 12 ring members” represents amonocyclic or bicyclic 5-membered to 12-membered heterocyclic ring thatis partly saturated or completely saturated. Examples of heteroatoms areN, O, and S. The heterocycle is unsubstituted or substituted on one ormore carbons or on one or more heteroatoms by identical or differentsubstituents. These substituents have been defined above for the radicalheteroaryl. In particular, the heterocyclic ring is monosubstituted orpolysubstituted, for example, monosubstituted, disubstituted,trisubstituted, or tetrasubstituted, on carbons by identical ordifferent groups selected from (C₁-C₈)-alkyl, for example,(C₁-C₄)-alkyl, (C₁-C₈)-alkoxy, for example, (C₁-C₄)-alkoxy such asmethoxy, phenyl-(C₁-C₄)-alkoxy, for example, benzyloxy, hydroxyl, oxo,halogen, nitro, amino, or trifluoromethyl, and/or it is substituted onthe ring nitrogens in the heterocyclic ring by (C₁-C₈)-alkyl, forexample, (C₁-C₄)-alkyl such as methyl or ethyl, by optionallysubstituted phenyl or phenyl-(C₁-C₄)-alkyl, for example, benzyl.Nitrogen heterocycles can also be present as N-oxides or as quaternarysalts.

Examples of the expressions heteroaryl having 5 to 14 ring members orheterocycle having 5 to 12 ring members are groups which are derivedfrom pyrrole, furan, thiophene, imidazole, pyrazole, oxazole, isoxazole,thiazole, isothiazole, tetrazole, 1,3,4-oxadiazole,1,2,3,5-oxathiadiazole-2-oxides, triazolones, oxadiazolones,isoxazolones, oxadiazolidinediones, triazoles which are substituted byF, CN, CF₃, or COO—(C₁-C₄)-alkyl, 3-hydroxypyrrole-2,4-diones,5-oxo-1,2,4-thiadiazoles, pyridine, pyrazine, pyrimidine, indole,isoindole, indazole, phthalazine, quinoline, isoquinoline, quinoxaline,quinazoline, cinnoline, carboline, and benzo-fused, cyclopenta-,cyclohexa-, or cyclohepta-fused derivatives of these hterocycles.Particularly preferred groups are 2- or 3-pyrrolyl, phenylpyrrolyl suchas 4- or 5-phenyl-2-pyrrolyl, 2-furyl, 2-thienyl, 4-imidazolyl,methylimidazolyl, for example, 1-methyl-2,4-, or 5-imidazolyl,1,3-thiazol-2-yl, 2-pyridyl, 3-pyridyl,4-pyridyl, 2-, 3-, or4-pyridyl-N-oxide, 2-pyrazinyl, 2-, 4-, or 5-pyrimidinyl, 2-, 3-, or5-indolyl, substituted 2-indolyl, for example, 1-methyl-, 5-methyl-,5-methoxy-, 5-benzyloxy-, 5-chloro-, or 4,5-dimethyl-2-indolyl,1-benzyl-2- or -3-indolyl, 4,5,6,7-tetrahydro-2-indolyl,cyclohepta[b]-5-pyrrolyl, 2-, 3-, or 4-quinolyl, 1-, 3-, or4-isoquinolyl, 1-oxo-1,2-dihydro-3-isoquinolyl, 2-quinoxalinyl,2-benzofuranyl, 2-benzothienyl, 2-benzoxazolyl, or benzothiazolyl, ordihydropyridinyl, pyrrolidinyl, for example, 2- or3-(N-methylpyrrolidinyl), piperazinyl, morpholinyl, thiomorpholinyl,tetrahydrothienyl, or benzodioxolanyl.

Thus methods and compositions are provided for the treatment of a hostwith a cellular proliferative disease, particularly a neoplasia. In thesubject methods, pharmaceutically acceptable imidazolines and anantiproliferative agent are administered, preferably systemically.

Methods and compositions are provided for the treatment of a host with acellular proliferative disease, particularly a neoplasia. In the subjectmethods, a pharmaceutically acceptable imidazoline is administered,preferably systemically, in conjunction with an antiproliferative agentto improve the anticancer effects. In a preferred embodiment, theimidazoline provides a chemopotentiator effect.

A chemical agent is a chemopotentiator when it enhances the effect of aknown antiproliferative drug in a more than additive fashion relative tothe activity of the chemopotentiator or antiproliferative agent usedalone. In some cases, a chemosensitizing effect may be observed. This isdefined as the effect of use of an agent that if used alone would notdemonstrate significant antitumor effects but would improve theantitumor effects of an antiproliferative agent in a more than additivefashion than the use of the antiproliferative agent by itself.

As used herein, the term imidazoline includes all members of thatchemical family including the forms and analogs thereof. The imidazolinefamily is defined by chemical structure as the ring structurespreviously described.

As used herein, antiproliferative agents are compounds, which inducecytostasis or cytotoxicity. Cytostasis is the inhibition of cells fromgrowing while cytotoxicity is defined as the killing of cells. Specificexamples of antiproliferative agents include: antimetabolites, such asmethotrexate, 5-fluorouracil, gemcitabine, cytarabine; anti-tubulinprotein agents such as the vinca alkaloids, paclitaxel, colchicine;hormone antagonists, such as tamoxifen, LHRH analogs; and nucleic aciddamaging agents such as the alkylating agents melphalan, BCNU, CCNU,thiotepa, intercalating agents such as doxorubicin and metalcoordination complexes such as cisplatin and carboplatin.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows the structures of compounds 1 to 20.

FIG. 2 shows the x-ray crystal structure of compound 1 which isrepresentative.

FIG. 3 is a EMSA of nuclear extracts with imidazolines 8 to 10. Lane 1,DNA only (control); lane 2, DNA, nuclear extract (10 μg) with p50homodimer (control); lane 3, DNA, nuclear extract (10 μg) with PMAactivation (control); lane 4, DNA, nuclear extract (10 μg) with no PMAactivation (control); lane 5, DNA, nuclear extract (10 μg) after PMAactivation with compound 8 (1.0 μM); lane 6, DNA, nuclear extract (10μg) after PMA activation with compound 8 (0.1 μM); lane 7, DNA, nuclearextract (10 μg) after PMA activation with compound 9 (1.0 μM); lane 8,DNA, nuclear extract (10 μg) after PMA activation with compound 9 (0.1μM); lane 9, DNA, nuclear extract (10 μg) after PMA activation withcompound 10 (0.1 μM); lane 10, DNA, nuclear extract (10 μg) after PMAactivation with compound 10 (0.1 μM).

FIGS. 4A and 4B show tumor growth delay with compounds 4 and 6.

DESCRIPTION OF PREFERRED EMBODIMENTS

A new class of imidazolines with anticancer (antitumor)anti-inflammatory activity and/or antimicrobial activity is described.Preferred compounds are shown in FIG. 1. The stereopositioning is shownin FIG. 2. The combination of these two key characteristics makes thisclass of imidazolines an extremely effective therapeutic drug to treatseptic shock as well as many other inflammatory disorders such as asthmaand infectious disorders. The objective of this invention is the use ofmulti-substituted imidazolines for therapeutic use as:

1) anti-inflammatory agents (for example in the treatment of asthma andrheumatoid arthritis).

2) antibacterial agents, including antiseptic agents.

The compounds of the present invention are very potent inhibitors ofNF-κB in vitro (less than 0.1 microMolar concentrations) and preliminaryexperiments in cells have indicated that the compounds are not cytotoxicover a 72 hour time period. Several of the imidazolines indicatedantimicrobial activity against several strains of bacteria with MIC's of50 microgram/milliliter.

The present invention also relates to the synthesis of the first classof imidazoline-type NF-κB inhibitors anti-inflammatory agents. Theimidazolines were prepared via a novel highly diastereoselectivemulticomponent synthesis using amino acid derived oxazolidinones asgeneral templates.

The general procedure for synthesis of Imidazoline-4-carboxylic acids isas follows: A solution of aldehyde (for example 0.57 mmol), amine (forexample 0.57 mmol) in dry CH₂Cl₂ (10 mL) was refluxed under N₂ for 2 h.A solution of the oxazolone (for example 0.57 mmol) in dry CH₂Cl₂ (forexample 5 mL) was added and the mixture was refluxed under N₂ for 6 hand then stirred overnight at room temperature. The product waspreferably either precipitated out from 1:1 CH₂Cl₂ or isolated aftersilica gel chromatography with 4:1 EtOAc/MeOH.

This is a novel highly diastereoselective multicomponent one-potsynthesis of aryl, acyl, alkyl and heterocyclic unsymmetricalsubstituted imidazolines. After screening a small number of Lewis acidsit was found that TMSCl (trimethylsilylchloride) promotes thecondensation of azlactones and imines to afford imidazolines in goodyields as single diastereometers (Scheme 1).

Acyl chlorides

where R is chiral can be used to obtain a single enantiomer. Theazlactones were prepared from different N-acyl-α-amino acids followed byEDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride)mediated dehydration to provide the pure azlactones in high yields(Schunk, S., et al., Organic letters 2, 907-910 (2000); and Sain, B., etal., Heterocycles 23 1611-1614 (1985)). The cycloaddition reactions withthe imines proceeded well at slightly elevated temperatures (for example40° C.) to provide the high substituted imidazolines in good yields. Theabsence of trimethylsilyl chloride resulted in the formation ofβ-lactams, presumably via a ketene intermediate (S. Peddibhotla, S.Jayakumar and J. J. Tepe, Highly Diastereoselective MulticomponentSynthesis of Unsymmetrical Imidazolines, Organic Letters 4, 3533-3535(2002)). Only the trans diastereomers of the imidazolines were observedin all of these reactions as determined by NOE experiments and X-raycrystallography. The diastereoselective multicomponent one-pot synthesisprovided a wide range of aryl, acyl, alkyl and heterocyclic substitutedimidazolines in excellent yields (Table 1).

TABLE 1

Yield compound R₁ R₂ R₃ R₄ R₅ (%)  1 Phenyl Methyl Phenyl Benzyl H 75  2Phenyl Methyl 4-meth- Benzyl H 78 oxyphenyl  3 Phenyl Methyl Phenyl4-Fluorophenyl H 74  4 Phenyl Phenyl Phenyl Benzyl H 65  5 Phenyl 1H-Phenyl Benzyl H 68 indol- 3-yl- methyl  6 Phenyl methyl pyridin-4-Benzyl H 76 yl  7^(a) Phenyl Methyl Phenyl H H 70  8 Phenyl MethylEthoxy- H H 72 carbonyl  9^(b) Phenyl Methyl pyridin-4- Benzyl Et 76 yl10^(c) Phenyl Methyl Phenyl Benzyl Et 75Table 1. Preparation of imidazolines 1-10. ^(a)After Hydrogenation (10%Pd/C, H₂ 1 atm) of compound 1, ^(b)After esterification (SOCl2, EtOH) ofcompound 6, ^(c)After esterification (SOCl2, EtOH) of compound 1.

While the complete mechanistic detail of this process is still underinvestigation, the reaction does not seem to proceed by activation ofthe carbonyl oxygen of the oxazolone by trimethylsilyl chloride, in turncausing ring-opening to the intermediate nitrilium ion as initiallyexpected (Ivanova, G. G., Tetrahedron 48 177-186 (1992)). Carrying outthe condensation in presence of slight excess of triethylamine haltedthe reaction altogether suggesting that acidic conditions were required.In addition, the addition of Lewis acids such as TiCl₄ or BF₃. OEt₂ didnot result in any product formation. In the light of these findings, itis proposed that the reaction probably proceeds by 1,3-dipolar type ofcycloaddition. Steric repulsion between the R₂ and R₃ moieties duringthe cycloaddition can explain the diastereoselectivity (Scheme 2).

EXAMPLES 1-20

Experimental Section:

1. Dl-(3S,4S)-1-Benzyl-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid SP-1-61

A solution of benzaldehyde (0.06 g, 0.57 mmol), benzylamine (0.061 g,0.57 mmol) in dry dichloromethane (15 mL) was refluxed under nitrogenfor 2 h. 2-Phenyl-4-methyl-4H-oxazolin-5-one (0.1 g, 0.57 mmol) andchlorotrimethylsilane (0.08 g, 0.74 mmol) were added and the mixture wasrefluxed under nitrogen for 6 h and then stirred overnight at roomtemperature. The reaction mixture was evaporated to dryness undervacuum. The product was precipitated our as a white solid using 1:1dichloromethane/hexanes mixture (0.155 g, 74%). ¹H NMR (300 MHz)(DMSO-d₆): δ 1.8 (3H, s), 4.05 (1H, d, J=15 Hz), 4.95 (1H, d, J=14.8Hz), 5.05 (1H, s), 7.05 (2H, s), 7.25-7.54 (8H, m), 7.74 (2H, t, J=7.2Hz), 7.83 (1H, t, J=6.9 Hz), 8.0 (2H, d, J=8.4 Hz); ¹³C NMR (75 MHz)(DMSO-d₆): δ 25.2, 48.8, 70.4, 73.3, 122.3, 127.8, 128.3, 128.5, 128.9,129.1, 129.3, 129.6, 129.7, 132.3, 133.2, 134, 166.1, 169.5; IR (neat):3350 cm⁻¹, 1738 cm¹; HRMS (EI): calculated for C₂₄H₂₂N₂O₂ [M−H]⁺369.1603, found [M−H]⁺ 369.1610; M.P.: decomposes at 185-190° C.

2. dl-(3S,4S)-1-Benzyl-5-(4-methoxyphenyl)-4-methyl-2-phenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid SP-1-63: A solution of p-anisaldehyde (0.077 g, 0.57 mmol),benzylamine (0.061 g, 0.57 mmol) in dry dichloromethane (15 mL) wasrefluxed under nitrogen for 2 h. 2-Phenyl-4-methyl-4H-oxazolin-5-one(0.1 g, 0.57 mmol) and chlorotrimethylsilane (0.08 g, 0.74 mmol) wereadded and the mixture was refluxed under nitrogen for 6 h and thenstirred overnight at room temperature. The reaction mixture wasevaporated to dryness under vacuum. The product was precipitated out asa white solid using 1:1 dichloromethane/hexanes mixture (0.180 g, 78%).¹H NMR (300 MHz) (CDCl₃+2 drops DMSO-d₆): δ 1.8 (3H, s), 3.8 (3H, s),3.95 (1H, d, J=15.3 Hz), 4.5 (1H, s), 4.9 (1H, d, J=15 Hz), 6.83-6.92(4H, m), 7.08-7.19 (3H, m), 7.3-7.4 (3H) dd, J₁=5.1 Hz, J₂=1.8 Hz),7.54-7.62 (2H, t, J=7.2 Hz), 762-7.68 (1H, t, J=7.2 Hz), 7.9 (2H, d,J=6.9 Hz); ¹³C NMR (75 MHz) (CD₃OD): δ 25.3, 48.8, 55.6, 70.9, 74.1,115.2, 122.2, 123, 125.5, 127.9, 128.4, 129.2, 129.3, 129.6, 129.9,132.8, 134.2, 161.1, 166.3, 168.4; IR (neat): 3388 cm⁻¹ 1738 cm⁻¹; HRMS(EI): calculated for C₂₅H₂₄N₂O₃ [M−H]⁺ 397.1709, found [M−H]⁺ 399.1717;M.P.: decomposes at 205-208° C.

3. dl-(3S,4S)-1-(4-Fluorophenyl)-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid SP-1-101: A solution of benzaldehyde (0.060 g, 0.57 mmol),4-fluoroaniline (0.063 g, 0.57 mmol) in dry dichloromethane (15 mL) wasrefluxed under nitrogen for 2 h. 2-Phenyl-4-methyl-4H-oxazolin-5-one(0.1 g, 0.57 mmol) and chlorotrimethylsilane (0.08 g, 0.74 mmol) wereadded and the mixture was refluxed under nitrogen for 6 h and thenstirred overnight at room temperature. The reaction mixture wasevaporated to dryness under vacuum. The product was precipitated out asa white solid using 1:1 dichloromethane/hexanes mixture (0.160 g, 74%).¹H NMR (300 MHz) (DMSO-d₆): δ 1.98 (3H, s), 5.98 (1H, s), 7.05-7.65(14H, m); ¹³C NMR (75 MHz) (DMSO-d₆) δ 25.2, 71.2, 77.9, 116.9, 117,117.1, 117.3, 123, 125.1, 125.3, 129.3, 129.4, 129.6, 130.1, 130.3,130.4, 130.5, 132.5, 133.3, 134.5, 160.4, 163.7, 165.3, 170.4; IR(neat): 3450 cm⁻¹, 1744 cm⁻¹. HRMS (EI): calculated for C₂₃H₁₉FN₂O₂[M−H]⁺ 373.1352, found [M−H]⁺ 373.1359; M.P.: decomposes at 230-232° C.

4. dl-(3S,4S)-1-Benzyl-2,4,5-triphenyl-4,5-dihydro-1H-imidazole-4-carboxylic acidSP-1-125: A solution of benzaldehyde (0.6 g, 5.7 mmol), benzylamine(0.61 g, 5.7 mmol) in dry dichloromethane (120 mL) was refluxed undernitrogen for 2 h. 2,4-Diphenyl-4H-oxazolin-5-one (1.35 g, 5.7 mmol) andchlorotrimethylsilane (0.8 g, 7.4 mmol) were added and the mixture wasrefluxed under nitrogen for 6 h and then stirred overnight at roomtemperature. The product was purified by silica-gel columnchromatography with 1:5 ethanol/ethyl acetate to afford 2.1 g of productin 65% yield as an off-white solid. ¹H NMR (300 MHz) (CDCL₃): δ 3.8 (1H,d, J=15.6 Hz), 4.62 (1H, d, J=15.6 Hz), 4.98 (1H, s), 6.58 (2H, d, J=8.1Hz), 7.05-7.65 (16H, m), 7.9 (2H, d, J=7.2 Hz); ¹³C NMR (75 MHz) (CDCl₆)δ 29.7, 48.3, 75.6, 79.1, 123.1, 125.7, 126.7, 127.3, 127.4, 127.9,128.1, 128.2, 128.8, 128.9, 129, 129.3, 132.9, 133.8, 136, 143.1, 164.8,168.1; IR (neat): 3400 cm⁻¹ (very broad), 1738 cm⁻¹; HRMS (EI):calculated for C₂₉H₂₄N₂O₂ [(M−H) CO₂]⁺ 387.1526 and observed[M−H)—CO₂]⁺387.1539; M. P.: decomposes at 153-155° C.

5. dl-(3S,4S)-1-Benzyl-4-(1H-indol-3-ylmethyl)-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid SP-1-128: A solution of benzaldehyde (0.6 g, 5.7 mmol), benzylamine(0.61 g, 5.7 mmol) in dry dichloromethane (120 mL) was refluxed undernitrogen for 2 h. 4-(1H-Indol-3-ylmethyl)-2-phenyl-4H-oxazol-5-one (1.65g, 5.7 mmol) and chlorotrimethylsilane (0.8 g, 7.4 mmol) were added andthe mixture was refluxed under nitrogen for 6 h and then stirredovernight at room temperature. The product was purified by silica-gelcolumn chromatography with 1:5 ethanol/ethyl acetate to afford 3.1 g ofproduct in 68% yield as an off-white solid. ¹H NMR (300 MHz) (DMSO-d₆):δ 3.95 (1H, d, J=16.2 Hz), 4.6 (1H, d, J=16.2 Hz), 5.25 (1H, s), 6.1(2H, d, J=7.8 Hz), 6.9-7.3 (5H, m), 7.3-8.0 (15H, m), ¹³C NMR (75 MHz)(DMSO-d₆) δ 169.6, 166, 136.5, 133.7, 132.5, 132.3, 129.7, 129.4, 128.9,128.7, 128.6, 127.9, 127.8, 126.7, 126.6, 122.7, 121.4, 119, 111, 105.8,74.4, 70.4, 48.5, 32.3; IR (neat): 3420 cm⁻¹ (very broad), 1741 cm⁻¹;HRMS(EI); calculated for C₃₂H₂₇N₃O₂ [M−H]⁺ 484.2025 and observed [M−H]⁺484.2011; M.P.: decomposes at >250° C.

6. dl-(3S,4S)-1-Benzyl-4-methyl-2-phenyl-5-pyridin-4yl-4,5-dihydro-1H-imidazole-4-carboxylicacid SP-1-150: A solution of pyridin-4-carboxalaldehyde (0.061 g, 0.57mmol), benzylamine (0.061 g, 0.57 mmol) in dry dichloromethane (15 mL)was refluxed under nitrogen for 2 h. 2-Phenyl-4-methyl 4H-oxazolin-5-one(0.1 g, 0.57 mmol) and chlorotrimethylsilane (0.08 g, 0.74 mmol) wereadded and the mixture was refluxed under nitrogen for 6 h and thenstirred overnight at room temperature. The reaction mixture wasevaporated to dryness under vacuum. The product was isolated using 4:1ethyl acetate/methanol as an off-white solid (0.161 g, 76%). ¹H NMR (300MHz) (DMSO-d₆): δ 1.8 (3H, s), 4.24 (1H, d, J=15.9 Hz), 4.9 (1H, d,J=14.8 Hz), 5.15 (1H, s), 7.0-7.15 (2H, m), 7.25-7.35 (3H, m), 7.45-7.5(2H, m), 7.7-7.9 (3H, m), 7.95-8.05 (2H, m), 8.6-8.7 (2H, m); ¹³C NMR(75 MHz) (DMSO-d₆) δ 25.1, 49.1, 70.6, 71.7, 122.1, 123, 127.9, 128.4,128.8, 129.2, 129.4, 132.8, 133.9, 141.4, 149.8, 166.5, 169.05; IR(neat); 3400 cm⁻¹, 1746 cm⁻¹; HRMS (EI): calculated for C₂₃H₂₁N₃O₂[M−H]⁺ 370.1556, found [M−H]⁺ 370.1556; M.P.: decomposes at 185-190° C.

7. d1 (3S,4S)-4-Methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylic acid:16/17 [JK1-1-135] To a well-stirred suspension ofimidazoline-4-carboxylic acid 10 (0.1 gm, 0.27 mmol) and cyclohexene(0.1 ml, 1.25 mmol) in dry THF (30 ml) added 10% Pd/C (45 mg, 0.06mmol). The suspension was refluxed for 36 h. The reaction mixture cooledto room temperature and ethanol (10 mL) was added. The mixture wasfiltered through a Celite bed, washed with ethanol and the filtrate wasevaporated under reduced pressure. The crude product was purified bycolumn silica-gel chromatography using ethanol, to yield a white solid(0.070 g, 93%). ¹H NMR (300 MHz) (DMSO-d₆) δ 1.76 (s, 3H), 5.34 (s, 1H),7.34-7.36 (b, 5H), 7.69 (dd, J=8.1, 7.2, 2H), 7.81 (1H, dd, J₁=6.9 Hzand J₂=7.2 Hz), 8.15 (2H, d, J=8.4 Hz); ¹³C NMR (75 MHz) (DMSO-d₆):25.32, 55.66, 70.79, 72.57, 123.12, 128.24, 128.96, 129.42, 129.67,130.12, 135.42, 136.24, 164.24, 170.77; IR (neat) 1734 cm⁻, 1616 cm⁻; MS(EI): calculated for C₁₇H₁₆N₂O₂ (m/z) 280.12 observed m/z: 280.1; M.P.:decomposes at 222-224° C.

8. dl-(3S,4S)-1-(4-Fluorophenyl)-4-methyl-2-phenyl-4,5-dihydro-1H-imidazole-4,5-dicarboxylicacid 5-ethyl ester SP-1-175: A solution of ethyl glyoxalate (0.058 g,0.57 mmol) as 50% solution in toluene (1.03 g/ml), 4-fluoroaniline(0.063 g, 0.57 mmol) in dry dichloromethane (15 mL) was refluxed undernitrogen for 2 h. 2-Phenyl-4-methyl-4H-oxazolin-5-one (0.1 g, 0.57 mmol)and chlorotrimethylsilane (0.08 g, 0.74 mmol) were added and the mixturewas refluxed under nitrogen for 6 h and then stirred overnight at roomtemperature. The reaction mixture was evaporated to dryness undervacuum. The product was purified by silica-gel column chromatographyusing 4:1 ethyl acetate/methanol, to yield a white solid (0.152 g, 72%).¹H NMR (300 MHz) (CD₃OD): δ 1.2 (3H, t, J=7.2 Hz), 2.03 (3H, s), 4.9(2H, dq, J₁=7.2 Hz, J₂=2.1 Hz), 5.48 (1H, s), 7.1-7.8 (9H, m); ¹³C NMR(75 MHz) (CD₃OD): δ 169.9, 166.2, 164.0, 162.1, 134.4, 131.5, 129.7,129.6, 129.5, 129.3, 121.8, 116.9, 116.7, 75.1, 69.1, 62.9, 24.2, 12.8;1R (neat): 3450 cm⁻¹, 1743 cm¹; HRMS (EI): calculated for C₂₀H₁₉FN₂O₄[M−H]⁺ 369.1251, and observed [M−H]⁺ 369.1255; M.P.: decomposes at190-193° C.

9. d1-(3S,4S)-1-Benzyl-4-methyl-2-phenyl-5-pyridin-4-yl-4,5-dihydro-1H-imidazole-4-carboxylicacid ethyl ester JK-1-183: To a well-stirred suspension of dl-(3S,4S)-1-Benzyl-4-methyl-2-phenyl-5-pyridin-4yl-4,5-dihydro-1H-imidazole-4-carboxylicacid 12 (0.1 g, 0.27 mmol) in dry dichloromethane (30 mL) at 0° C. addeda solution of oxallyl chloride (0.14 g, 1.1 mmol) in dry dichloromethane(5 mL). A solution of DMF (0.001 mL) was added to the reaction mixtureand was stirred at 0° C. for another 2 h. The dichloromethane wasevaporated under vacuum and the reaction mixture cooled to 0° C. afterwhich absolute ethanol (20 mL) was added. The solution was allowed tostir for an additional 1 h. The solvent was evaporated under vacuum andthe reaction mixture diluted with dichloromethane (30 mL) and washedwith saturated sodium bicarbonate (1-0 mL) and water (10 mL). Theorganic layer was dried over sodium sulfate and was concentrated undervacuum to yield crude product, which was further purified by silica-gelcolumn chromatography using ethyl acetate, to yield a pale yellow oil(0.097 gm, 91%). ¹H NMR (300 MHz) (CDCl₃): δ 0.86 (3H, t, J=7.2 Hz),1.57 (3H, s), 3.64 (2H, q, J=7.2 Hz), 3.83 (1H, d, J=15.3 Hz), 4.27 (1H,s), 4.77 (1H, d, J=15.3, Hz), 6.97 (2H, dd, J₁=7.2 Hz and J₂=2.4 Hz),7.22-7.54 (6H, m), 7.31-7.54 (2H, m), 7.78-7.81 (2H, m), 8.59-8.61 (2H,m). ¹³C NMR (75 MHz) (CDCl₃): δ 13.45, 27.13, 49.47, 60.83, 71.87,77.94, 122.56, 127.79, 127.93, 128.55, 128.70, 130.21, 130.51, 135.82,146.59, 149.75, 166.02, 171.37; IR (neat): 1734 cm⁻¹; MS (EI):calculated for C₂₅H₂₆N₂O₂ (m/z) 399.19 observed m/z: 399.3.

10. d1-(3S,4S)-1-Benzyl-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid ethyl ester JK-1-186: To a well-stirred suspension ofImidazoline-4-carboxylic acid 10 (0.1 gm, 0.27 mmol) in dry methylenechloride (30 ml) at 0° C. added a solution of oxallyl chloride (0.14 g,1.1 mmol) in dry dichloromethane (5 ml). A solution of DMF (0.001 mL) indry dichloromethane (1 mL) was added to the reaction mixture and wasstirred at 0° C. for another 2 h. The dichloromethane was evaporatedunder vacuum and the reaction mixture cooled to 0° C. after whichabsolute ethanol (20 ml) was added. The solution was allowed to stir foran additional 1 h. The solvent was evaporated under vacuum and thereaction mixture diluted with dichloromethane (30 ml) and washed withsaturated sodium bicarbonate (10 ml) and water (10 ml). The organiclayer was dried over sodium sulfate and was concentrated under vacuum toyield crude product, which was further purified by silica-gel columnchromatography using ethyl acetate, to yield colorless oil (0.095 gm,89%). ¹H NMR (300 MHz, CDCl₃): δ 0.84 (3H, t, J=7.2 Hz), 1.57 (3H, s),3.60 (2H, q, J=7.2 Hz), 3.85 (1H, d, J=15.3 Hz), 4.32 (1H, s), 4.74 (1H,d, J=15.3 Hz), 6.98 (2H, dd, J₁=6.9 Hz and J₂=2.1 Hz), 7.27-7.35 (m,8H), 7.49-7.51 (2H, m), 7.76-7.79 (2H, m); ¹³C NMR (75 MHz, CDCl₃): δ13.80, 27.13, 49.12, 60.06, 71.31, 127.98, 128.03, 128.12, 128.67,129.02, 129.11, 130.96, 136.40, 136.80, 166.11, 171.78; IR (neat); 1730cm⁻¹, 1495 cm⁻¹; MS (EI): calculated for C₂₆H₂₆N₂O₂ (m/z) 398.2 observedm/z=398.9.

11. dl-(3S,4S)-1-Methoxycarbonylmethyl-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid JK-1-199: To a well stirred solution of2-Phenyl-4-methyl-4H-oxazolin-5-one (0.5 g, 2.85 mmol) and TMSCl (0.37g, 3.42 mmol) in dry dichloromethane (50 ml) added a solution of(Benzylidene-amino)-acetic acid methyl ester (0. gm, mmol) in drymethylene chloride (20 ml) and the mixture was refluxed under nitrogenfor 10 h and then stirred overnight at room temperature. The reactionmixture was evaporated to dryness under vacuum. The product wasprecipitated out as a white solid using a 1:1 dichloromethane/hexanesmixture (0.70 g, 70%). ¹H NMR (300 MHz) (CD₃OD): δ 1.99 (3H, (1H, d,J=18.3 Hz), 4.53 (1H, d, J=18.3 Hz), 5.39 (1H, s), 7.47-7.50 (5H, m),7.74-7.87 (5H, m). ¹³C NMR (75 MHz) (CD₃OD): δ 24.23, 52.09, 70.83,75.38, 121.84, 128.26, 128.69, 129.52, 129.75, 131.78, 134.02, 167.59,168.62, 169.19; IR (neat): 3468 cm⁻¹, 1747 cm⁻¹; MS (EI): calculated forC₂₀H₂₀N₂O₄ (m/z) 352.14 observed m/z=353.2; M.P.: decomposes at 215-217°C. s), 3.67 (3H, s), 3.96.

12.1-Benzyl-5-(4-methoxy-phenyl)-2,4-dimethyl-4,5-dihydro-1H-imidazole-4-carboxylicacid SP-1-189: A solution of p-anisaldehyde (1.4 g, 10.4 mmol),benzylamine (1.11 g, 10.4 mmol) in dry dichloromethane (150 mL) wasrefluxed under nitrogen for 2 h. 2,4-dimethyl-4H-oxazolin-5-one SP-1-188(1f) (1 g, 8.7 mmol) and chlorotrimethylsilane (1.22 g, 11.3 mmol) wereadded and the mixture was refluxed under nitrogen for 6 h and thenstirred overnight at room temperature. The reaction mixture wasevaporated to dryness under vacuum. The product was precipitated out asa white solid using a 1:1 dichloromethane/hexanes mixture (1.9 g, 65%).¹H NMR (300 MHz) (CDCl₃): δ 1.13 (3H, s), 2.43 (3H, s), 3.83 (3H, s),4.17 (1H, d, J=15.9 Hz), 4.57 (1H, d, J=15.9 Hz), 5.8 (1H, s) 6.92 (2H,d, J=8 Hz), 7.05 (2H, d, J=8 Hz) 7.2-7.4 (5H, m); ¹³C NMR (75 MHz)(CDCl₃): δ 12.3, 21.9, 47.8, 55.2, 70.4, 114.3, 125.2, 126.9, 128.5,129.3, 133.3, 159.9, 163.2, 174.8; IR (neat): 3388 cm⁻¹; 1738 cm⁻¹; HRMS(EI): calculated for C₂₀H₂₂N₂O₃ [M−H]⁺ (m/z)=337.1552, found (m/z)337.1548.

13. dl-(3S,4S)-1-(2-Ethoxycarbonyl-ethyl)-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid JK-1-215: To a well stirred solution of2-Phenyl-4-dimethyl-4H-oxazolin-5-one (1.0 g, 5.7 mmol) and TMSCl (1 ml,6.8 mmol) in dry dichloromethane (80 ml) added a solution of3-(Benzylidene-amiono)-propionic acid ethyl ester (1.4 gm, 6.8 mmol) indry methylene chloride (60 ml) and the mixture was refluxed undernitrogen for 10 h and then stirred overnight at room temperature. Thereaction mixture was evaporoated to dryness under vacuum. The productwas precipitated out as a white solid using a 1:1dichloromethane/hexanes mixture (1.08 g, 51.4%). ¹H NMR (500 MHz)(CD₃OD): δ 1.17 (t, J=7.5, 3H), 1.9 (s, 3H), 2.47-2.52 (m, 1H),2.52-2.71 (m, 1H), 3.34-3.39 (m, 1H), 3.40-4.09 (m, 3H), 5.42 (s, 1H),7.46-7.49 (m, 5H), 7.72-7.87 (m, 5H); ¹³C NMR (100 MHz) (CD₃OD): δ13.35, 24.87, 30.64, 41.64, 61.00, 70.94, 73.51, 122.77, 128.99, 129.21,129.80, 130.10, 132.78, 134.09, 167.32, 169.81, 170.9. IR (neat): 3481cm⁻¹, 1743 cm⁻¹; MS (EI): calculated for C₂₂H₂₄N₂O₄ (m/z) 380.44observed m/z=380.7. M.P.: decomposes at 218-220° C.

14. dl-(3S,4S)-1-(1-Methoxycarbonyl-ethyl)-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid JK-1-192: To a well stirred solution of2-Phenyl-4-methyl-4H-oxazolin-5-one (0.25 g, 1.5 mmol) and TMSCl (0.23ml, 1.8 mmol) in dry dichloromethane (50 ml) added a solution of2-(Benzlidene-amino)-propionic acid methyl ester (0.34 gm, 1.8 mmol) indry methylene chloride (20 ml) and the mixture was refluxed undernitrogen for 10 h and then stirred overnight at room temperature. Thereaction mixture was evaporated to dryness under vacuum. The product wasprecipitated out as a white solid using a 1:1 dichloromethane/hexanesmixture (0.340 g, 66%). ¹H NMR (300 MHz) (CD₃OD): δ 1.19 (d, J=6.9, 3H),2.06 (s, 3H), 3.38 (s, 3H), 4.89 (q, J=6.9, 1H), 544 (s, 1H), 7.43-7.46(5H, m), 7.75-7.85 (5H, m). ¹³C NMR (75 MHz) (CD₃OD): δ 14.9, 25.6,52.7, 56.7, 71.9, 72.5, 122.2, 128.8, 128.9, 129.6, 130.0, 134.5, 135.8,169.2, 169.4, 170.4, IR (neat): 3431 cm⁻¹, 1740 cm⁻¹; MS (EI):calculated for C₂₁H₂₂N₂O₄ (m/z) 366.4 observed m/z=366.6. M.P.:decomposes at 222-226° C.

15.1-Benzyl-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazol-4-yl)-methanol 14[JK-1-123]: To a well stirred suspension of Lithium aluminum hydride(0.12 gm, 0.3 mmol) in dry THF (5 ml) added a solution of1-Benzyl-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid (0.1 gm, 0.27 mmol) in dry THF (5 ml) at 0° C. drop wise, stirredat same temperature for 15 min quenched with ice cold saturated ammoniumchloride solution [Caution: Ammonium chloride solution kept at 0° C. forabout 30 min.; and should be added with extreme care; highly exothermicreaction and the reaction mixture should be at 0° C.] then added about10 ml of 10% HCl. The reaction mixture diluted with excess of ethylacetate (100 ml) washed with water (20 ml) dried over anhydrous sodiumsulfate, filtered through a fluted filter paper and the organic layerevaporated under reduced pressure to yield the crude product which waspurified by column chromatography using ethyl acetate. Yield: 79%;viscous oil, IR (neat): 3314, 2928, 1643, 1516; δ H (300 MHz, CD₃Cl₃): δ1.25 (s, 3H), 3.48 (d, J=12, 1H), 3.56 (d, J=11.8, 1H), 3.75 (d, 12.9,1H), 3.87 (s, 1H), 3.94 (d, J=12.9, 1H), 7.28-7.54 (m, 13H), 7.77-7.79(m, 2H), 8.06 (brs, 1H); δ C (75 MHz, CDCl₃): δ 17.25, 51.67, 61.54,66.28, 66.93, 127.266, 127.68, 128.26, 128.56, 128.82, 129.06, 131.77,135.48 138.03, 139.90, 167.91; m/z: 357.2.

16.1-Benzyl-4-(2-methoxycarbonyl-ethyl)-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid SP-1-201: A solution of benzaldehyde (0.252 g, 2.4 mmol),benzylamine (0.258 g, 2.4 mmol) in dry dichloromethane (100 mL) wasrefluxed under nitrogen for 2 h.3-(5-Oxo-2-phenyl-4,5-dihydro-oxazol-4-yl)-propionic acid methyl esterSP-1-182 (1e)(0.5 g, 2 mmol) and chlorotrimethylsilane (0.282 g, 2.6mmol) were added and the mixture was refluxed under nitrogen for 6 h andthen stirred overnight at room temperature. The reaction mixture wasevaporated to dryness under vacuum. The product was precipitated out asa white solid using a 1:1 dichloromethane/hexanes mixture (0.54 g, 60%).¹H NMR (300 MHz) (CDCl₃): δ 2.05-2.25 (2H, m), 2.3-2.5 (2H, m), 3.55(3H, s), 4.38 (2H, ddd, J₁=4 Hz, J₂=9 Hz, J₃=25 Hz), 4.86 (1H, q,J=3.3), 7.1-7.6 (12H, m), 7.7-7.9 (4H, m); ¹³C NMR (75 MHz) (CDCl₃): δ27.6, 30.1, 43.3, 51.6, 52.7, 127.1, 127.2, 127.3, 128.2, 128.3, 131.5,131.6, 133.3, 137.8, 167.5, 171.4, 173.6; IR (neat): 1734 cm⁻¹, 1653cm⁻¹; MS (EI): calculated for C₂₄H₂₂N₂O₂ (m/z) 442.5, found (m/z) 443.

17. dl-(3S,4S)-1-Benzyl-2,4-dimethyl-5-phenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid: 15[JK-1-238]. To a well stirred solution of2,4-dimethyl-4H-oxazolin-5-one (0.4 g, 3.5 mmol) and TMSCl (0.58 ml, 4.2mmol) in dry dichloromethane (60 ml) added a solution ofBenzyl-benzylidene-amine (0.82 gm, 4.2 mmol) in dry methylene chloride(40 ml) and the mixture was refluxed under nitrogen for 10 h and thenstirred overnight at room temperature. The reaction mixture wasevaporated to dryness under vacuum. The product was precipitated out asa white solid using a 1:1 dichloromethane/hexanes mixture (0.60 g, 60%).¹H NMR (300 MHz) (CD₃OD): δ 1.11 (s, 3H), 2.47 (s, 3H), 4.17 (d, J=16.2,1H), 4.63 (q, J=16.2, 1H), 5.84 (s, 1H), 7.04-7.07 (m, 2H), 7.27-7.42(m, 7H). ¹³C NMR (75 MHz) (CD₃OD): δ 12.62, 22.12, 48.27, 70.39, 71.25,127.31, 128.83, 129.28, 129.58, 133.40, 133.46, 164.12, 175.19. IR(neat): 3431 cm⁻¹, 1740 cm⁻¹; MS (EI); calculated for C₁₉H₂₀N₂O₂ (m/z)308.37 observed m/z=308.3, M.P.; decomposes at 232-234° C.

18. dl-(3S,4S)-1-Benzyl-2,4-diphenyl-5-pyridin-4-yl-4,5-dihydro-1H-imidazole-4-carboxylicacid SP-1-195: A solution of pyridin-4-carboxylaldehyde (0.61 g, 0.57mmol), benzylamine (0.61 g, 5.7 mmol) in dry dichloromethane (120 mL)was refluxed under nitrogen for 2 h. 2,4-Diphenyl-4H-oxazolin-5-one(1.35 g, 5.7 mmol) and chlorotrimethylsilane (0.8 g, 7.4 mmol) wereadded and the mixture was refluxed under nitrogen for 6 h and thenstirred overnight at room temperature. The product was purified byprecipitation from dichloromethane/ether mixture to afford 1.35 g of theproduct in 55% yield as an off-white solid. ¹H NMR (300 MHz) (CDCl₃): δ4 (1H, d, J=15.6 Hz), 5.0 (1H, d, J=15.6 Hz), 5.38 (1H, s), 7.1-7.65(17H, m), 8.5 (2H, d, J=7.2 Hz); ¹³C NMR (75 MHz) (CDCl₃): δ 45.2, 66.3,75.6, 123.7, 126.5, 126.9, 128.5, 128.6, 128.8, 129.2, 129.3, 131.9,133.5, 134.4, 136.2, 143.4, 149.7, 166.6, 166.9; IR (neat): 3400 cm⁻¹(very broad), 1733 cm⁻¹; MS (EI): calculated for C₂₄H₂₂N₂O₂ (m/z)434.34, found (m/z) 434.2.

Compounds 19 and 20 Synthesis of1-Benzyl-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid (1-phenyl-ethyl)-amide from1-Benzyl-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid: JK-1-309

To a well-stirred suspension of1-Benzyl-4-methyl-2,5-diphenyl-4,5-dihydro-1H-imidazole-4-carboxylicacid (1.0 g, 0.27 mmol) in dry methylene chloride (25 ml),(S)-(−)-1-Phenyl-ethylamine (0.36 g, 29 mmol) was added EDCIHCl (0.57 g,29 mmol), after five minutes added a solution of DMAP (0.35 gm, 29 mmol)in methylene chloride (10 ml) and stirred for 5-6 hrs. The reactionmixture was washed with water (2×10 ml), saturated sodium bicarbonate(20 ml), water (20 ml), 2N HCl (20 ml) and then with water (30 ml). Theorganic layer dried over sodium sulfate and evaporated under reducedpressure. The crude product was purified by column silica-gelchromatography using ethyl acetate hexane mixture (1:1).

19: Yield (0.26 g, 40.7%). {[α]_(D)=+41.5°)} ¹H NMR (300 MHz): δ 1.02(d, J=6.9, 3H), 1.56 (s, 3H), 3.85 (d, J=15.6, 1H), 4.40 (s, 1H), 4.66(d, J=15.6, 1H), 4.72 (t, J=6.9, 1H), 7.07-7.09 (m, 2H), 7.17-7.55 (m,16H), 7.69-7.73 (m, 2H); ¹³C NMR (75 MHz): 21.39, 27.56, 48.09, 48.73,72.66, 126.52, 127.24, 127.71, 127.99, 128.42, 128.57, 128.67, 128.95,129.01, 129.14, 130.75, 130.82, 137.38, 137.60, 143.29, 165.44, 171.61.

20: (0.24 g, 38%). {[α]_(D)=37.7°)} ¹H NMR (300 MHz): δ 1.40 (d, J=7.23H), 1.61 (s, 3H), 3.77 (d, J=15.6, 1H), 4.37 (s, 1H), 4.60 (d, J=15.6,1H), 4.75 (t, J=7.5, 1H), 6.922-7.090 (m, 2H), 7.11-7.22 (m, 13H),7.507-7.529 (m, 3H), 7.651-7.682 (m, 2H): ¹³C NMR (75 MHz): 21.58,28.08, 47.97, 48.59, 72.62, 126.66, 126.99, 127.200, 127.69, 127.96,128.21, 128.51, 128.58, 128.64, 129.13, 129.122, 130.70, 130.83,137.184, 137.22, 143.28, 165.35, 171.62.

EXAMPLE 11

All compounds were evaluated for their potential anti-inflammatoryactivity by examining the activity of NF-κB in vitro in nuclear extractsusing the procedure from Breton and Charbot-Fletcher (Breton, J. J., etal., J. Pharmacol Exp Ther 282 459-466 (1997)). Briefly, Human Jurkatleukemia T-cells (clone E6-1; Amer. Type Culture Collection, Rockville,Md.) are grown in RPMI-1640 Media (Gibco-BRL, Rockville, Md.)supplemented with 10% Fetal Bovine Serum, Penicillin (614 ηg/mL),Streptomycin (10 μg/mL) and Hepes Buffer, pH 7.2 at 37° C., 5% CO₂. TheJurkat cells (1×10⁷ cells/mL) are subsequently treated with variousconcentrations of imidazoline for 30 min. at 37° C. followed by PMAstimulation (5.0 ng/mL) for an additional 5 hours. Nuclear extracts areincubated for 20 minutes with a double stranded Cy3 labeled NF-κBconsensus oligonucleotide, 5′-AGTTGAGGGGACTTTCCCAGGC-3′ at roomtemperature. The crude mixture is loaded on a 5% non-denaturingpolyacrylamide gel prepared in 1× Tris borate/EDTA buffer andelectrophoresed at 200 V for 2 hours. After electrophoresis the gel isanalyzed using a phosphorimager (Biorad FX plus) for detection of theNF-κB-DNA binding.

Treatment of the cells to the imidazolines exhibited a significantinhibition of nuclear NF-κB activity. FIG. 3 clearly illustrates adecrease of nuclear NF-κB-DNA binding by imidazolines 8 to 10 (FIG. 3,lanes 5-10).

Cells treated with the imidazolines exhibited a significant inhibitionof nuclear NF-κB activity (FIG. 3). FIG. 3 clearly illustrates asignificant decrease of nuclear NF-κB-DNA binding in the presence 100 nMconcentration of imidazolines 8-10 (FIG. 3, lanes 5-10).

The apparent absence of a slow moving band in lane 5 is indicative ofsignificant (94%) NF-κB inhibition by compound 8 at 1 micromolarconcentration in Jurkat Leukemia T-cells. Lane 6 indicates 88%inhibition of NF-κB-DNA binding in the nucleus by 100 nanomolarconcentrations of compound 8.

EXAMPLE 12

All compounds were tested for their ability to inhibit NF-κB and thecollected data is shown in Table 2. Currently, the most active compoundin the series is the heterocyclic imidazoline 9 which exhibited 88%inhibition of NF-κB at 100 nM concentrations. Preliminary resultsindicate that the imidazolines do not exhibit significant cytotoxicityfor up to 72 hours.

TABLE 2 compound concentration % inhibition 1   1 μM 19% 2   1 μM 68% 3  1 μM 35% 4   1 μM 65% 5   1 μM  0% 6 0.1 μM 84% 7 0.1 μM 38% 8 0.1 μM88% 9 0.1 μM 71% 10  0.1 μM 22%Table 2. Inhibition of NF-κB by imidazolines 1-10. The most activecompound in this series was compound 8.

IC₅₀ values in Mammalian Jurkat cells Leukemia T cells: IC₅₀ value isdefined as the concentration of compounds at which 50% of theprotein/enzyme is inhibited in cells.

TABLE 3 Compound 1. IC₅₀ = 1.95 micromolar Compound 2. IC₅₀ = 40nanomolar Compound 3. IC₅₀ = 6.5 nanomolar Compound 4. IC₅₀ = 73nanomolar Compound 5. IC₅₀ = Not tested Compound 6. IC₅₀ = 0.3micromolar Compound 7. IC₅₀ = 20 nanomolar

EXAMPLE 13

Compounds 4, 6 and 7 were tested for the inhibition of bacteria. A totalof 9 bacterial strains were screened. The following Gram-negative andGram-positive bacteria were included: Staphylococcus aureus,Enterobacter aerogenes, Esherichia coli, Klebsiella pneumonia,Pseudomonas aeruginosa, Serratia marcescens, Bacillus cerius, Bacillussubtillus and micrococcus luteus. Bacterial isolates were removed fromstorage, streaked on to nutrient agar plates and incubated for 18-24hours at 35° C. A working bacterial suspension was prepared bysuspending 3-5 isolated colonies in 5 mL saline solution. The turbidityof this suspension was carefully adjusted photometrically to equal thatof a 0.5 McFarland standard. The zone diameters were determined by astandardized disk diffusion method using cation-supplementedMueller-Hinton agar according to NCCLS guidelines (National Committeefor Clinical Laboratory Standards. Methods for dilution AntimicrobialSusceptibility Tests for Bacteria that Grow Aerobically. Fifth Edition:Approved Standard M7-A5. Wayne, Pa: NCCLS (2000)). Minimum inhibitoryconcentrations (MICs) were considered the lowest concentration that gavea clear zone of inhibition. The inoculated agar plates were incubatedfor 16-20 hours at 35° C. in ambient air. The diameters of the zoneswere read in millimeters. The results are shown in Table 4.

TABLE 4 Microbe MIC Compound 4 Bacillus subtillus 13 mm 50 μg Bacilluscereus 11 mm 50 μg Micrococcus luteus 12 mm 200 μg Staphylococcus aureus12 mm 200 μg Compound 6 Micrococcus luteus 10 mm 200 ηg Compound 7Micrococcus luteus 10 mm 56 ηg

EXAMPLE 14

Treatment of Imidazoline in RIF-1 Murine Tumor Model

Several of the NF-κB inhibitors (compounds 1, 3, 4 and 6) were tested inanimals. Tumor cells were injected, bilaterally, into the backs of mice.When tumors reached 100 mm³, the mice were treated with anintraperitoneal injection of the compound. Tumor volumes were measured 3times a week until they reached 4 times the size they were on the firsttreatment day. Data is recorded as “Days to 4×’ or ratio of ‘Days to 4×”of the treated over untreated controls.

Combinational treatment of the mice with cis-platin (CDDP) andcamptothecin (CPT) in the presence of compound 4 (1-SP-4-84) exhibitedconsiderable chemopotentiation of cis-platin (FIG. 4A). In addition,this group had 4 of the 8 tumors that remained <4× its volume at day 22of the experiment. No significant chemopotentiation of camptothecin inthe presence of compound 4 was shown.

Compound 6 (1-SP-6-95) exhibited significant chemopotentiation ofcis-platin as well as camptothecin (FIG. 4B). However, chemopotentiationby 6 was not as pronounced as seen with compound 4.

Combinational treatment of the mice with cis-platin (CDDP) andcamptothecin (CPT) in the presence and absence of the imidazolinesindicated that compounds 1 and 3 showed no significant chemopotentiationof either cis-platin or camptothecin (data not shown).

Combinational therapy of compound 4 with cis-platin showed a tumorgrowth delay (in days) of more than 10.26 days as compared to cis-platin(0.82 days) or camptothecin (3.79 days) alone (Table 5). In addition,half of the tumors in this RIF-1 murine model did not reach the 4× tumorvolume cut-off point at day 22 days when exposed to combinationaltreatment with compound 4.

TABLE 5 Antitumor efficacy of imidazolines as measured by the RIF-1murine model of tumor growth delay. CGX-E060 # of Dose Days to 4x DaysTreatment Tumors Route (mg/kg) (Ave ± SE) T/C Median Delay Untreated 10— — 7.3 ± 0.6 0.0 7.0 0.00 Cis-platin 8 IP  4 8.1 ± 0.4 1.1 7.8 0.82Compound 1 8 IP 100 6.5 ± 0.3 0.9 6.4 −0.57 Compound 3 8 IP 100 7.6 ±1.0 1.0 6.8 −0.20 Compound 4 6/8 IP 100 6.4 ± 0.2 0.9 6.5 −0.56 Compound6 8 IP 100 6.6 ± 0.3 0.9 6.6 −0.41 CDDP + 1 8 IP 4/100 8.8 ± 0.5 1.2 9.12.05 CDDP + 3 8 IP 4/100 8.8 ± 0.3 1.2 8.5 1.49 CDDP + 4* 8 IP4/100 >17.1 ± 1.9  >2.3 >17.3 >10.26 CDDP + 6 8 IP 4/100 9.8 ± 0.3 1.310.1 3.09 Camptothecin 8 IP  6 10.3 ± 0.6  1.4 10.8 3.79 CPT + 1 8 IP6/100 10.2 ± 0.4  1.4 10.3 3.27 CPT + 3 8 IP 6/100 8.8 ± 0.6 1.2 8.71.70 CPT + 4 4/8 IP 6/100 10.8 ± 0.4  1.5 11.1 4.07 CPT + 6 8 IP 6/10011.8 ± 0.9  1.6 10.7 3.72 *This group had 4 of 8 tumors < 4x at Day 22.Abbreviations: CDDP (cis-platin) and CPT (camptothecin) and IP(intraperitoneal injection).

This data illustrates the efficacy of the imidazolines in thechemopotentiation of commonly used anticancer drugs. Inhibition ofchemoresistance by these novel NF-κB inhibitors (especially compound 4)results in a significant delay of tumor growth as compared to treatmentof the tumors with the anticancer drug alone.

In pharmaceutical compositions, the imidazoline is inhibitory at adosage of 1 to 1,000 micrograms per milliliter or gram. It can be usedin a ratio of 1 to 100 or 100 to 1 with the antitumor compound. In apreferred embodiment, one or more of the imidazolines for treating apatient are provided to the patient at an inhibitory dose in apharmaceutically acceptable carrier. As such, the imidazolines areprocessed with pharmaceutical carrier substances by methods well knownin the art such as by means of conventional mixing, granulating,coating, suspending and encapsulating methods, into the customarypreparations for oral or rectal administration. Thus, imidazolinepreparations for oral application can be obtained by combining one ormore of the anthraquinones with solid pharmaceutical carriers;optionally granulating the resulting mixture; and processing the mixtureor granulate, if desired and/or optionally after the addition ofsuitable auxiliaries, into the form of tablets or dragee cores.

Suitable pharmaceutical carriers for solid preparations are, inparticular, fillers such as sugar, for example, lactose, saccharose,mannitol or sorbitol, cellulose preparations and/or calcium phosphates,for example, tricalcium phosphate or calcium hydrogen phosphate; alsobinding agents, such as starch paste, with the use, for example, ofmaize, wheat, rice or potato starch, gelatine, tragacanth, methylcellulose, hydroxypropylmethyl cellulose, sodium carboxymethyl celluloseand/or polyvinylpyrrolidone, esters of polyacrylates orpolymethacrylates with partially free functional groups; and/or, ifrequired, effervescent agents, such as the above-mentioned starches,also carboxymethyl starch, cross-linked polyvinylpyrrolidone, agar, oralginic acid or a salt thereof, such as sodium alginate. Auxiliaries areprimarily flow-regulating agents and lubricating agents, for example,silicic acid, talcum, stearic acid or salts thereof, such as magnesiumstearate or calcium stearate. Dragee cores are provided with suitablecoatings, optionally resistant to gastric juices, whereby there areused, inter alia, concentrated sugar solutions optionally containing gumarabic, talcum, polyvinylpyrrolidone, and/or titanium dioxide, lacquersolutions in aqueous solvents or, for producing coatings resistant tostomach juices, solutions of esters of polyacrylates orpolymethacrylates having partially free functional groups, or ofsuitable cellulose preparations such as acetylcellulose phthalate orhydroxypropyl-methylcellulose phthalate, with or without suitablesofteners such as phthalic acid ester or triacetin. Dyestuffs orpigments may be added to the tablets or dragee coatings, for example foridentification or marking of the various doses of active ingredient.

Imidazoline preparations comprising one or more of the anthraquinoneswhich can be administered orally further include hard gelatine capsules,as well as hard or soft closed capsules made from gelatine and, ifrequired, a softener such as glycerin or sorbitol. The hard gelatinecapsules can contain one or more of the imidazolines in the form of agranulate, for example in admixture with fillers such as maize starch,optionally granulated wheat starch, binders or lubricants such astalcum, magnesium stearate or colloidal silicic acid, and optionallystabilizers. In closed capsules, the one or more of the imidazolines isin the form of a powder or granulate; or it is preferably present in theform of a suspension in suitable solvent, whereby for stabilizing thesuspensions there can be added, for example, glycerin monostearate.

Other imidazoline preparations to be administered orally are, forexample, aqueous suspensions prepared in the usual manner, whichsuspensions contain the one or more of the anthraquinones in thesuspended form and at a concentration rendering a single dosesufficient. The aqueous suspensions either contain at most small amountsof stabilizers and/or flavoring substances, for example, sweeteningagents such as saccharin-sodium, or as syrups contain a certain amountof sugar and/or sorbitol or similar substances. Also suitable are, forexample, concentrates or concentrated suspensions for the preparation ofshakes. Such concentrates can also be packed in single-dose amounts.

Suitable imidazoline preparations for rectal administration are, forexample, suppositories consisting of a mixture of one or more of theimidazolines with a suppository foundation substance. Such substancesare, in particular, natural or synthetic triglyceride mixtures. Alsosuitable are gelatine rectal capsules consisting of a suspension of theone or more of the imidazolines in a foundation substance. Suitablefoundation substances are, for example, liquid triglycerides, of higheror, in particular, medium saturated fatty acids.

Likewise of particular interest are preparations containing the finelyground one or more of the imidazolines, preferably that having a medianof particle size of 5 μm or less, in admixture with a starch, especiallywith maize starch or wheat starch, also, for example, with potato starchor rice starch. They are produced preferably by means of a brief mixingin a high-speed mixer having a propeller-like, sharp-edged stirringdevice, for example with a mixing time of between 3 and 10 minutes, andin the case of larger amounts of constituents with cooling if necessary.In this mixing process, the particles of the one or more of theimidazolines are uniformly deposited, with a continuing reduction of thesize of some particles, onto the starch particles. The mixturesmentioned can be processed with the customary, for example, theaforementioned, auxiliaries into the form of solid dosage units; i.e.,pressed for example into the form of tablets or dragees or filled intocapsules. They can however also be used directly, or after the additionof auxiliaries, for example, pharmaceutically acceptable wetting agentsand distributing agents, such as esters of polyoxyethylene sorbitanswith higher fatty acids or sodium lauryl sulphate, and/or flavoringsubstances, as concentrates for the preparation of aqueous suspensions,for example, with about 5- to 20-fold amount of water. Instead ofcombining the imidazoline/starch mixture with a surface-active substanceor with other auxiliaries, these substances may also be added to thewater used to prepare the suspension. The concentrates for producingsuspensions, consisting of the one or more of the imidazoline/starchmixtures and optionally auxiliaries, can be packed in single-doseamounts, if required in an airtight and moisture-proof manner.

In addition, the one or more imidazolines can be administered to apatient intraperitoneally, intranasally, subcutaneously, orintravenously. In general, for intraperitoneal, intranasal,subcutaneous, or intravenous administration, one or more of theimidazolines are provided by dissolving, suspending or emulsifying themin an aqueous or nonaqueous solvent, such as vegetable or other similaroils, synthetic aliphatic acid glycerides, esters of higher aliphaticacids or propylene glycol; and if desired, with conventional additivessuch as solubilizers, isotonic agents, suspending agents, emulsifyingagents, stabilizers and preservatives. Preferably, the one or moreimidazolines are provided in a composition acceptable forintraperitoneal, subcutaneous, or intravenous use in warm-bloodedanimals or humans. For example, such compositions can comprise aphysiologically acceptable solution such as a buffered phosphate saltsolution as a carrier for the one or more anthraquinones. Preferably,the solution is at a physiological pH. In particular embodiments, thecomposition is injected directly into the patient perfused through thetumor by intravenous administration.

Preparations according to the present invention comprise one or more ofthe imidazolines at a concentration suitable for administration towarm-blooded animals or humans which concentration is, depending on themode of administration, between about 0.3% and 95%, preferably betweenabout 2.5% and 90%. In the case of suspensions, the concentration isusually not higher than 30%, preferably about 2.5%; and conversely inthe case of tablets, dragees and capsules with the one or more of theanthraquinones, the concentration is preferably not lower than about0.3%, in order to ensure an easy ingestion of the required doses of theone or more imidazolines. The treatment of patients with thepreparations comprising one or more of the imidazolines is carried outpreferably by one or more administrations of a dose of the one or moreimidazoline which over time is sufficient to substantially inhibitNF-κB. If required, the doses can be administered daily or divided intoseveral partial doses which are administered at intervals of severalhours. In particular cases, the preparations can be used in conjunctionwith or following one or more other therapies such as radiation orchemotherapy. The administered dose of the one or more imidazolines isdependent both on the patient (species of warm-blooded animal or human)to be treated, the general condition of the patient to be treated, andon the type of disease to be treated.

It is intended that the foregoing description be only illustrative ofthe present invention and that the present invention be limited only bythe hereinafter appended claims.

1. A method for inhibiting inflammation in a mammal in need thereofwhich comprises administering an imidazoline of the formula:

to the mammal in an amount sufficient to inhibit the inflammation. 2.The method of claim 1 wherein the mammal is human.
 3. The method ofclaim 1 wherein the mammal is a non-human mammal.
 4. The method of anyone of claims 1, 2 or 3 wherein the administration is orally to themammal.
 5. The method of any one of claims 1, 2 or 3 wherein theadministration is topically to the mammal.
 6. The method of any one ofclaims 1, 2 or 3 wherein the administration is by injection into themammal.
 7. The method of any one of claims 1, 2 or 3 wherein theadministration is intravenous into the mammal.